CICS-UBI - Health Sciences Research Center, University of Beira Interior, Av Infante D Henrique, 6200-506 Covilhã, Portugal.
Int J Biochem Cell Biol. 2012 Nov;44(11):2077-84. doi: 10.1016/j.biocel.2012.08.013. Epub 2012 Aug 16.
Prostate cancer (PCa) progresses from an early stage, confined to prostate, to a more aggressive metastasized cancer related with loss of androgen responsiveness. Although, it has been recognized that PCa cells have unique metabolic features, their glycolytic profile in androgen-dependent and androgen-independent stages of disease is much less known. Hence, the main purpose of this study was to compare glucose metabolism in androgen-responsive (LNCaP) and androgen-nonresponsive (PC3) PCa cells. Cell culture medium was collected and differences in glucose consumption and, lactate and alanine production were measured using Proton Nuclear Magnetic Resonance ((1)H NMR) spectra analysis. The mRNA and protein expression of glucose transporters (GLUT1 and GLUT3), phosphofructokinase 1 (PFK1), lactate dehydrogenase (LDH) and monocarboxylate transporter (MCT4) were determined by real-time PCR and Western Blot, respectively. The obtained results demonstrate that androgen-responsive (LNCaP) and androgen-nonresponsive (PC3) cells consumed similar amounts of glucose, whereas PC3 cells present higher lactate production. This increase in lactate production was concomitant with higher levels of MCT4 protein, increased LDH activity and higher lactate/alanine ratio, also suggesting increased levels of oxidative stress in PC3 cells. However, protein levels of LDH, associated with lactate metabolism, and GLUT3, involved in glucose uptake, were decreased in PC3 comparatively with LNCaP. Androgen-responsive and nonresponsive PCa cells present distinct glycolytic metabolism profiles, which suggest that targeting LDH and MCT4 metabolic pathways may be an important step for the development of new diagnostic and therapeutic strategies in the different stages of PCa.
前列腺癌(PCa)从早期局限于前列腺的阶段发展为更具侵袭性的转移性癌症,与雄激素反应性丧失有关。尽管已经认识到 PCa 细胞具有独特的代谢特征,但它们在疾病的雄激素依赖性和雄激素非依赖性阶段的糖酵解谱知之甚少。因此,本研究的主要目的是比较雄激素反应性(LNCaP)和雄激素非反应性(PC3)PCa 细胞的葡萄糖代谢。收集细胞培养物上清液,使用质子磁共振(1H NMR)谱分析测量葡萄糖消耗以及乳酸和丙氨酸的产生差异。通过实时 PCR 和 Western Blot 分别测定葡萄糖转运蛋白(GLUT1 和 GLUT3)、磷酸果糖激酶 1(PFK1)、乳酸脱氢酶(LDH)和单羧酸转运蛋白(MCT4)的 mRNA 和蛋白表达。结果表明,雄激素反应性(LNCaP)和雄激素非反应性(PC3)细胞消耗的葡萄糖量相似,而 PC3 细胞表现出更高的乳酸产量。这种乳酸产量的增加与 MCT4 蛋白水平升高、LDH 活性增加以及乳酸/丙氨酸比值升高同时发生,也表明 PC3 细胞中氧化应激水平升高。然而,与 LNCaP 相比,PC3 中与乳酸代谢相关的 LDH 相关蛋白和参与葡萄糖摄取的 GLUT3 蛋白水平降低。雄激素反应性和非反应性 PCa 细胞表现出不同的糖酵解代谢谱,这表明靶向 LDH 和 MCT4 代谢途径可能是开发新的诊断和治疗策略在 PCa 的不同阶段的重要步骤。